mouse aortic vsmcs  (ATCC)

Journal: Redox Biology

Article Title: Ultrasmall Cu 2−x Se nanoparticles alleviate vascular calcification through inhibiting oxidative stress and NF-κB/NLRP3-mediated inflammation

doi: 10.1016/j.redox.2025.103961

Figure Lengend Snippet: CSP NPs inhibit VSMC calcification via downregulating NLRP3 signaling pathway. Rat vascular smooth muscle cells (VSMCs) (A–D), human VSMCs (E–F), or rat aortic rings (G–H) were treated with growth medium (GM), calcifying medium (CM), or CM with Nig (5 μM) in the absence or presence of CSP NPs (10 μg/mL) for 7 days (n = 4). (A) Representative alizarin red staining images of VSMCs. Scale bar = 500 μm. (B) Quantitative analysis of alizarin red staining was assessed using a microplate reader. (C–D) Runx2 and BMP2 expression was analyzed by Western blot and quantified by densitometry. (E) Representative alizarin red staining images of human VSMCs. Scale bar = 500 μm. (F) Quantitative analysis of alizarin red staining was assessed using a microplate reader. (G) Representative alizarin red staining images of rat aortic rings. Scale bar = 500 μm (low power) and 250 μm (high power). (H) Quantitative analysis of alizarin red positive area by ImageJ software. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Human VSMCs (CRL-1999, ATCC, USA) were cultured under identical conditions.

Techniques: Staining, Expressing, Western Blot, Software

Journal: bioRxiv

Article Title: miR-18a-5p upregulates Orai1 expression to promote vascular smooth muscle cell proliferation and neointimal hyperplasia after injury

doi: 10.64898/2026.01.20.700592

Figure Lengend Snippet: A, B. Bar graphs showing levels of Orai1 (A) and SARAF (B) mRNA in A7r5 cells transfected with mimic of miR18a-5p mimic (miR-18a-5p) and scrambled miRNA (Control). miRNA expression was calculated using the 2 −ΔΔCt method normalized to 18S expression. C. Relative Luciferase Assay upon transfection with pGL3 Basic 3Kb Orai1 and scrambled miRNA (Control), and pGL3 Basic 3Kb Orai1 and miR18a-5p mimic (miR-18a-5p) in A7r5 cells. D. Representative immunoblots (top) and summary data (bottom) showing the protein expression of Orai1 normalized to its corresponding α-SMA in A7r5 cells transfected with scrambled miRNA (Control) or miR18a-5p mimic (miR 18a-5p). Representative traces (E) and mean values (F) of thapsigargin-induced Ca 2+ influx in Fura-2 loaded A7r5 cells. Recordings were acquired from A7r5 cells transfected with scrambled miRNA (Control) (n = 193), or miR-18a-5p mimic (miR-18a-5p) (n = 179), and preincubated with 2 µM of TG in Ca 2+ free solution for 3 minutes. GSK 7975A (GSK,10 μM) was applied to inhibit SOCE. F. Δratio indicate the difference between the peak ratio after extracellular Ca 2+ addition and its level immediately before the addition. (n = 4 cell culture). Values are presented as mean ± SD. (*), (**), and (****) indicate significance with p < 0.05, p < 0.01, and p < 0.0001, respectively.

Article Snippet: In the case of rat aortic A7r5 VSMC (CRL-1444, ATCC, Manassas, VA, US), they were expanded in basal media (DMEM supplemented with 10% FBS) and 100 U/ml penicillin and streptomycin.

Techniques: Transfection, Control, Expressing, Luciferase, Western Blot, Cell Culture

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Gin A inhibits HG-induced proliferation of A10 cells (A) Structural formula of Gingerenone A (Gin A). (B) Effect of high glucose (HG) on the proliferation of A10 cells. A10 cells were incubated with 10-, 15-, 30-, or 50-mM glucose for 24 h. Cell proliferation was assessed by MTT assay (n = 8, * P < 0.05 vs. control). (C,D) Effect of Gin A on the proliferation of A10 cells. A10 cells were cultured for 24 h in normal medium or HG (30 mM) with Gin A at 0.1, 1, 10 or 100 μM. Cell proliferation was evaluated by MTT assay (C) , and automated cell counting (D) (n = 8, * P < 0.05 vs. control). (E) Representative PCNA immunofluorescence images of A10 cells cultured under control, HG (30 mM) and HG plus Gin A (10 μM) conditions for 24 h. Staining was repeated in three independent experiments with similar results; images are shown for qualitative illustration only and were not subjected to statistical analysis. (F) PCNA protein levels in A10 cells after 24 h of treatment with control medium, Gin A (10 μM), HG (30 mM) or HG plus Gin A (10 μM) for 24 h. Representative western blots and quantification of PCNA expression quantification of PCNA expression (normalized to H3) are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. HG alone).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Incubation, MTT Assay, Control, Cell Culture, Cell Counting, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Gin A reduces HG-induced migration of A10 cells (A) Representative Transwell images showing A10 cell migration after 24 h of culture under control conditions, Gin A (10 μM), HG (30 mM) or HG plus Gin A (10 μM). (B) Representative wound-healing images at 0 and 24 h for A10 cells treated as in (A) . (C) Effect of Gin A on the increase in the number of migrating cells induced by HG (n = 10; * P < 0.05 vs. control; # P < 0.05 vs. HG alone). (D) Effect of Gin A on the increased migration distance induced by HG (n = 10; * P < 0.05 vs. control; # P < 0.05 vs. HG alone).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Migration, Control

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Gin A attenuates HG-induced oxidative stress in A10 cells (A) Intracellular ROS levels in A10 cells cultured for 24 h in control medium or HG (30 mM) in the absence or presence of Gin A (10 μM). ROS was evaluated by DHE staining and quantitative statistical data (n = 10; * P < 0.05 vs. control; # P < 0.05 vs. HG alone). (B–D) Oxidative stress-related indicators in A10 cells treated with control medium, Gin A alone (10 μM), HG (30 mM) or HG plus Gin A (10 μM) for 24 h. MDA (B) , T-AOC (C) and SOD activity (D) measured using assay kits (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. HG alone). (E) NOX4 protein expression in A10 cells cultured for 24 h under control, HG or HG plus Gin A conditions. Representative western blots and NOX4/GAPDH ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. HG alone).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Cell Culture, Control, Staining, Activity Assay, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Gin A activates AMPK and suppresses mTOR/S6K1 signaling in A10 cells (A) Concentration-dependent effects of Gin A on AMPK phosphorylation. A10 cells were treated with Gin A at 0.1, 1, 10 or 100 μM for 24 h, and p-AMPK/AMPK ratios were determined by Western blot. (B) Time course of AMPK activation by Gin A. Cells were exposed to Gin A (10 μM) for 0.5, 1, 3, 6 or 24 h, and p-AMPK/AMPK levels were determined by Western blot. (C) Effects of Gin A on mTOR phosphorylation. A10 cells were treated with Gin A at 0.1, 1, 10 or 100 μM for 24 h, and p-mTOR/mTOR ratios were determined by Western blot. (D) Effects of Gin A on S6K1 phosphorylation. A10 cells were treated with Gin A at 0.1, 1, 10 or 100 μM for 24 h, and p-S6K1/S6K1 ratios determined by Western blot (n = 6; * P < 0.05 vs. control).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Concentration Assay, Phospho-proteomics, Western Blot, Activation Assay, Control

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Role of AMPK activation in the Gin A-mediated inhibition of the proliferation and migration of HG-treated A10 cells (A) Effect of the AMPK inhibitor Compound C on Gin A-induced AMPK activation in HG-treated A10 cells. Cells were pre-incubated with Compound C for 1 h and then exposed to HG (30 mM) with or without Gin A (10 μM) for 24 h. Representative blots and p-AMPK/AMPK ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. Gin A alone). (B–D) Effects of Compound C on the Gin A-induced inhibition of A10 cell proliferation and migration. Cell proliferation was determined by the MTT assay (B) . Cell migration was determined by Transwell (C) and wound healing (D) assays (n = 8; * P < 0.05 vs. control; # P < 0.05 vs. HG alone; & P < 0.05 vs. HG + Gin A). (E,F) Effects of Compound C on Gin A-mediated inhibition of mTOR/S6K1 signaling in HG-treated A10 cells. Representative blots and quantification of p-mTOR/mTOR (E) and p-S6K1/S6K1 (F) ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. HG alone; & P < 0.05 vs. HG + Gin A).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Activation Assay, Inhibition, Migration, Incubation, Control, MTT Assay